Antioxidant Mixture Research Articles

Influence of sub-lethal antioxidant doses, water potential and temperature on growth, sclerotia, aflatoxins and aflD (=nor-1) expression by Aspergillus flavus RCP08108

Effects of interacting conditions of sub-lethal levels of antioxidants, water potential (Ψ) and temperature were evaluated on growth, sclerotial characteristics, aflatoxin B1 (AFB1) production and aflD (=nor-1) gene expression by Aspergillus flavus strain RCP08108. These studies were carried out on peanut meal extract agar osmotically modified to −2.8,−7.1, −9.9 and −16.0MPa and incubated at 28 and 20°C. The food grade antioxidants added were butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) at (1+1mM−M1) and (5+5mM−M2). To relate the aflD expression after toxigenic A. flavus grew under interacting stress conditions, real-time PCR was used. Antioxidant mixtures caused a higher and significant (p<0.001) reduction in growth rate. The major impact on size and volume sclerotia was produced by Ψ; followed by antioxidant mixtures. High AFB1 levels were observed in response to the M1 applied at −7.1MPa. Induction of the aflD gene was observed in response to the M1 treatment at −2.8, −7.1 and −9.9MPa; but significant decreases of AFB1 production and aflD transcripts were observed; when the fungus grew in the presence of the M2 treatment.These results showed that it is necessary to apply food-grade antioxidants into the peanut storage system at levels higher than 5mM. This is an important tool to avoid sub-lethal antioxidant doses that can lead to fungal growth, increase resistance structures, and stimulate aflD gene expression and AFB1 accumulation in this substrate.

Particularities of corrective action of polar lipids and bioantioxidants from sea hydrobionts at imbalances of lipid and carbohydrate metabolism

A total mixture of phospho- and glycolipids from sea macrophytes Sargassum pallidum, Ulva fenestrata, Zostera marina was separated and the fatty acid composition was determined. Biological activity of the mixtures of polar lipids and natural antioxidants echinochrome A from flat sea urchin Scaphechinus mirabilis and polyphenolic complex from sea grass Zostera marina was studied in rats with experimental model of atherosclerosis and diabetes. These experiments revealed optimal compositions for mixtures of polar lipids and antioxidants, which possess high medical-corrective activity. Proposed mechanisms of action of the polar lipids (containing different polyunsaturated fatty acids) and antioxidants studied are presented. These compositions may be used for creation of new biologically-active additives and drugs.

Resolution of phenolic antioxidant mixtures employing a voltammetric bio-electronic tongue

This work reports the application of a Bio-Electronic Tongue (BioET) system made from an array of enzymatic biosensors in the analysis of polyphenols, focusing on major polyphenols found in wine. For this, the biosensor array was formed by a set of epoxy-graphite biosensors, bulk-modified with different redox enzymes (tyrosinase and laccase) and copper nanoparticles, aimed at the simultaneous determination of the different polyphenols. Departure information was the set of voltammograms generated with the biosensor array, selecting some characteristic features in order to reduce the data for the Artificial Neural Network (ANN). Finally, after the ANN model optimization, it was used for the resolution and quantification of each compound. Catechol, caffeic acid and catechin formed the three-analyte case study resolved in this work. Good prediction ability was attained, therefore allowing the separate quantification of the three phenols with predicted vs. expected slope better than 0.970 for the external test set (n = 10). Finally, BioET has been also tested with spiked wine samples with good recovery yields (values of 104%, 117% and 122% for catechol, caffeic acid and catechin, respectively).

It is Time to Formulate an Antioxidant Mixture for Management of Diabetes and its Complications: Notice for Pharmaceutical Industries

It is Time to Formulate an Antioxidant Mixture for Management of Diabetes and its Complications: Notice for Pharmaceutical Industries

Implementation of response surface methodology to assess the antiradical behaviour in mixtures of ascorbic acid and α-tocopherol with grape (Vitis vinifera) stem extracts

The efficiency of grape stem extracts to express antiradical activity was assessed using two different approaches and DPPH as the radical probe. In addition, the mixture effects when the extracts were combined with ascorbic acid (AA) and α-tocopherol (α-Tcp) were also evaluated. The approaches included a simple linear regression analysis between the response (antiradical activity) and concentration, but also a response surface methodology, which permitted the monitoring of the response upon simultaneous variation of both the concentration of the total polyphenols of the extracts and either of the antioxidants (AA and α-Tcp). The deployment of linear regression poses important constraints with regard to concentration ranges, whereas response surface methodology might be a valuable statistical tool for similar assessments and credible modelling of binary mixtures of antioxidants. In all combinations tested it was found that an antagonism was manifested, presumably as a result of AA and α-Tcp regeneration by the extract polyphenols, at the expense of the latter.

DSC study of stabilizing effect of antioxidant mixtures in styrene-butadiene rubber

The type of cooperation between antioxidants in the binary mixtures of four substituted diphenylamines and phenotiazine in the stabilization of styrene-butadiene rubber has been tested. Thermooxidation of the samples has been studied by differential scanning calorimetry under non-isothermal conditions. The protection factors of the individual stabilizers and their mixtures were determined. The synergy factors were applied to asses the type of cooperation of antioxidants in the mixtures. From their values it can be concluded that the type of cooperation depends on temperature. The highest synergistic effect has been observed for the mixture of phenotiazine and [4-(1-methyl-1-phenyl-ethyl)-phenyl]-phenylamine.

Oxidative stabilization of ultra-high omega-3 concentrates as ethyl esters or triacylglycerols

A comparative study of the oxidative stability of “ultra-high” ω3 concentrates (80%) from fish oils as triacylglycerols (TAG) or ethyl esters (EE), as well as their stabilization by supercritical extracts of rosemary (SER), α-tocopherol or their mixture, by Rancimat method and throughout storage time, was carried out. No significant differences were found between EE and TAG on oxidative stability when measured by Rancimat conditions in the range of 50−90°C. However, storage experiments revealed a poor stability of the EE concentrate form, measured by the monitorization of peroxide and p-anisidine values. Concerning the stabilization by antioxidants, there was a lack of antioxidant effect of evaluated compounds when Rancimat assays were performed at 70°C, whereas an antioxidant effect began to evidence at 60°C, and a clearest antioxidant protection was measured at 50°C for the mixture of the SER plus α-tocopherol in both EE and TAG forms. This selected binary mixture efficiently stabilized the ω3-TAG concentrate throughout 50days of storage conditions, whereas the stabilization of ω3-EE concentrate was worse. Therefore, the combination of SER and α-tocopherol seemed to be a good antioxidant mixture for the efficient stabilization of extremely labile ultra-high ω3 concentrates, especially in the form of ω3 TAG oils.

Dietary antioxidant supplements: Benefits of their combined use

Several dietary supplements claim medicinal benefits due to their composition in hydrophilic and lipophilic molecules, natural extracts or synthetic compounds with antioxidant properties. In the present work, the antioxidant activity of selected supplements taken in pills, capsules or infusions were studied either individually or combined. Linear discriminant analysis (LDA) was used to categorize the condensed formulations (pills and capsules), infusion bags and combined samples according with their antioxidant activity measured by radical scavenging activity, reducing power and lipid peroxidation inhibition using brain homogenates as models. AAF proved to have the highest antioxidant activity in all the assayed methods, either singly taken or included in mixtures. Furthermore, the mixtures containing this supplement revealed synergistic effects in 92% of the cases. The intake of antioxidant mixtures might provide some additional benefits.

Investigation of antioxidant synergisms and antagonisms among thymol, carvacrol, thymoquinone and p-cymene in a model system using the Briggs–Rauscher oscillating reaction

Auto-oxidation of lipids has been recognised as a major deterioration process affecting both the sensory and nutritional quality of foods; the application of antioxidants is one of the technically simplest ways of reducing this process. Previous reports showed that these compounds were found in selected aromatic plants and that thymol, carvacrol, thymoquinone and p-cymene as constituents of different essential oils possess substantial antioxidant activity. In the present study their antioxidant capacities were evaluated using the Briggs–Rauscher oscillating reaction method.Our results indicate that the majority of the antioxidant mixtures demonstrated some level of discrepancy in antioxidant capacity when compared to individual values of their constituents. The strong antagonistic effect was found only when thymoquinone was paired with p-cymene. All other combinations of two antioxidants clearly showed some degree of synergistic effect. The mixtures of three antioxidants showed a decrease in antioxidant capacity when compared to their individual values. The results obtained for more complex mixtures of four antioxidants showed a decrease in antioxidant capacity, indicating and confirming that the antagonistic effect between thymoquinone and p-cymene overrides the synergistic effect of other pairs.

Effect of Catechins Extract of Green Tea on Quality Protection of Black Tea-bag During Storage

Green tea is one of the richest sources of antioxidants especially catechins, which are studied most because of their health benefits. These valuable compounds may lose their activity and oxidize to thearobigin during fermentation stage or black tea processing. The main objective of this research is applying antioxidant mixtures extracted from green tea on packaging material of black tea and determining the amount of antioxidants released from the tea bags by their immersion into boiling water. The green tea extracts prepared by a two-step extraction process in water at 50°C and 80°C in four individual time periods, i.e., 10min, 10min; 10min, 20min; 20min, 10min; 20min, 20min (1). Final extract from each process was analyzed by HPLC and DPPH tests and then sprayed on the tea bag packaging material.The activity and concentration of antioxidants (catechins) released from treated tea bags were investigated immediately and after 3 months storage. Sensory properties of the tea beverage samples were also studied. The results (Tables 1, 2) show that the concentration of antioxidants increases by 66–76% as compared to control i.e., untreated black tea. Tea beverage samples were more astringent as compared to untreated black tea. Under different time/temperature conditions the best results were obtained by the sample extracted by a two-step process (20min at 50°C) and (10min at 80°C) with the highest concentration of antioxidants, lowest astringency and best stability during 3 months storage.

Influence of cryoprotectants glycerol and amides, combined with antioxidants on quality of frozen-thawed boar sperm

The present study was undertaken to examine whether the cooling and freezing extenders containing a mixture of antioxidants (AOs) catalase, Na-pyruvate and mercaptoethanol and one of three types of cryoprotectants (CPs) would be able to improve the quality of frozen-thawed boar sperm. The collected semen, only the sperm-rich fraction, was diluted 1:1 with Androhep plus™ extender, stored at 15 °C for 2 h and centrifuged. The centrifuged sperm pellet was re-suspended in lactose-egg yolk extender and divided into four groups for mixing with freezing extenders containing different kinds of CPs at 5 °C: (I) glycerol (GLY) as control; (II) GLY with AOs; (III) dimethyl formamide (DMF) with AOs and (IV) dimethyl acetamide (DMA) with AOs. Processed sperm were packaged in 0.25-mL straws and frozen using a controlled rate freezer. After thawing, the diluted thawed sperm were incubated at 38 °C for 10 min and was assessed for motility by CASA, membrane/acrosome integrity by FITC-PNA/PI and DNA integrity (DFI) by SCSA. All sperm parameters evaluated, except DFI, were negatively affected ( P < 0.001) when using DMF (III) or DMA (IV) as CPs instead of GLY (I and II). Total sperm motility was lower ( P < 0.001) in the samples supplemented with AOs (32.4 ± 1.2, 23.9 ± 1.5, 6.9 ± 0.7, and 10.3 ± 0.9%, for treatments I, II, III and IV, respectively). The quality of sperm frozen in DMF was not different from DMA ( P > 0.05). There was no difference in DFI among the studied groups ( P > 0.05). In conclusion, based on the present results, addition of AOs to cooling and freezing extenders and/or replacement of GLY with DMF or DMA could not improve quality of frozen-thawed boar sperm.

Methodology of the spectrophotometric analysis of organic mixtures: Application of statistical models

Deviations of the absorbance of mixtures from additivity can be detected and predicted using statistical models obtained in the complete factorial experiment. The models correlate the deviations with the composition of a mixture; this allows the optimization of the procedures of spectrophotometric analysis. The advantages and limitations of this approach are demonstrated on an example of the analysis of pharmaceutical preparations and antioxidant mixtures.

Peculiarities of the corrective effects of polar lipids and bioantioxidants from sea hydrobionts in impairments of lipid and carbohydrate metabolism

A total mixture of phospho- and glycolipids from sea macrophytes Sargassum pallidum, Ulva fenestrata, Zostera marinawas separated and the fatty acid composition was determined. Biological activity of the mixtures of polar lipids and natural antioxidants (echinochrome A from the flat sea urchin Scaphechinus mirabilis and a polyphenolic complex from the sea grass Zostera marina) were studied under conditions of impairments of carbohydrate and lipid metabolism. Doses and compositions of mixtures of polar lipids and antioxidants possessing high corrective activity were optimized in mice with the experimental model of atherosclerosis and diabetes. Based on these results possible mechanisms of the effects of polar lipids containing various polyunsaturated fatty acids and the investigated antioxidants have been proposed. The developed compositions may be used for creation of new biologically active additives and drugs.

Bioactive antioxidant mixtures promote proliferation and migration on human oral fibroblasts

ObjectiveAntioxidants (AOs) are the first line of defence against free radical damage and are critical for maintaining optimum health and well being. The need for AOs becomes even more critical with increased exposure to free radicals generated by pollution, cigarette smoke, drugs, illness, stress and exercise. Antioxidant supplementation is an excellent way of improving free radical protection. The aim of this study was to provide cytotoxicity, proliferation and migration data on the in vitro effects of bioactive AO mixtures on human oral fibroblasts. MethodsHuman oral fibroblasts were obtained from human gingival (HGF) and periodontal (HPDL) tissues. Each of these oral fibroblasts was cultured separately in three concentrations of the bioactive pure polyphenol and turmeric derivative mixtures; resveratrol (R), ferulic acid (F), phloretin (P) and tetrahydrocurcuminoids (T); [(RFT), (PFR), and (PFT)]. Cell viability, proliferation, morphology and migratory behaviour were analysed in vitro using high throughput in vitro 96 well plate wound assay. ResultsRFT decreased (10−3M) and increased (10−5M) cell number in HGF cells. Three concentrations (10−3, 10−4, and 10−5M) of PFR and PFT increased DNA synthesis in HGF cells. PFT promoted cell migration but PFR and RFT had no significant change in HGF wound healing rates in a 96 well plate assay monolayer wound. In the HPDL cells, the 10−4M concentration of both RFT and PFT increased cell number at 72h and 96h whereas the lower concentration 10−5M of RFT significantly stimulated cell number at 96h. PFR (10−3M and 10−5M) and PFT (10−3M) increased DNA synthesis after 48h treatment in HPDL cells. ConclusionsHigh and low concentrations (10−3–10−5M) of these AOs (RFT, PFR) may have beneficial effects on functional mechanisms regulating fibroblast migration and proliferation during gingival healing or periodontal repair.

A new potent natural antioxidant mixture provides global protection against oxidative skin cell damage

Oxidative stress occurs when there is an over production of free radicals and cells are not able to neutralize them by their own antioxidant mechanisms. These excess of free radicals will attack cellular macromolecules leading to cell damage, function impairment or death. Because of that, antioxidant substances have been largely used in products to offer complementary protection. In this study a new mixture of three known antioxidants (cocoa, green tea and alpha-tocopherol) was evaluated and its antioxidant protection was assessed focusing on its capacity to protect main cell macromolecules. Results have shown that it has a high antioxidant capacity by protecting lipids, DNA and proteins against oxidative damage. The antioxidant effect of the mixture on cells was also investigated and it was able to reduce oxidative stress generated by lipopolisacharide in human fibroblasts. Finally, as the mixture has proved to be highly antioxidant, its effect on cell senescence was evaluated, and it was demonstrated that fibroblasts in culture had delayed senescence when treated with these actives on a mixture. All results together provide important data about a new antioxidant mixture that uses a small amount of actives and is able to protect cell against oxidative damages in a global way.

Qualitative and quantitative electroanalysis of synthetic phenolic antioxidant mixtures in edible oils based on their acid–base properties

A simple electroanalytical method using square wave voltammetry at a Pt band ultramicroelectrode to perform a qualitative and quantitative analysis of different synthetic antioxidant mixtures permitted by official regulations in edible oils is proposed. The methodology was based on the comparison of voltammetric signals obtained in acetonitrile+0.1M (C4H9)4NF6P with those recorded in the same reaction medium when different aliquots of (C4H9)4NOH were added to allow a qualitative differentiation between antioxidants. Firstly, studies on solutions prepared from commercial reagents were carried out. Then, the results obtained were transferred to the analysis of a real matrix, i.e., an edible olive oil. From real samples spiked with a known amount of different synthetic antioxidant mixtures, we could deduce the presence of these antioxidants by comparing results obtained in the neutral medium with those obtained after the successive addition of base. The standard addition method was used to quantify the individually spiked synthetic antioxidants in the real sample. Recovery percentages were between 88% and 118%. The reproducibility was 1.5%, 3.1%, 4.1% and 4.1% in ACN+0.1M TBAHFP and 1.5%, 4.6%, 6.6% and 2.5% in Bz/EtOH (1:2)+0.1M H2SO4 for TBHQ, BHA, BHT and PG, respectively. The repeatability was 1% for PG in both media. These parameters show a good system performance.

Effects of 6 weeks of n-3 fatty acids and antioxidant mixture on lipid peroxidation at rest and postexercise

The purpose of this randomized study was to measure the influence of 6weeks of LCPUFA (600mg EPA and 400mg DHA per day) supplementation alone or in association with 30mg vitamin E, 60mg vitamin C and 6mg β-carotene on resting and exercise-induced lipid peroxidation in judoists (n=36). Blood samples were collected at rest before (T (1)) and after the supplementation period, in preexercise (T (2)) and postexercise (T (3)) conditions, for analysis of α-tocopherol, retinol, lag phase (Lp) before free radical-induced oxidation, maximum rate of oxidation (R (max)) during the propagating chain reaction, maximum amount of conjugated dienes (CD(max)) accumulated after the propagation phase, and nitric oxide, malondialdehyde and lipoperoxide (POOL) concentrations. Dietary data were collected using a 7-day diet record. There were no significant differences among treatment groups with respect to habitual intakes of energy from fat, carbohydrate, or protein. At T (1), there were no significant differences among treatment groups with respect to lipid peroxidation, lag phase, and levels of α-tocopherol or retinol. The consumption of an n-3 LC PUFA supplement increased oxidative stress at rest and did not attenuate the exercise-induced oxidative stress. The addition of antioxidants did not prevent the formation of oxidation products at rest. On the contrary, it seems that the combination of antioxidants added to the n-3 LCPUFA supplement led to a decrease in, CD(max), R (max), and POOL and MDA concentrations after a judo training session.

Antioxidant/prooxidant properties of model phenolic compounds: Part I. Studies on equimolar mixtures by chemiluminescence and cyclic voltammetry

The electrochemical behaviour and the antioxidant effectiveness of ascorbic acid (AA), caffeic acid (CA), quercetin (Q), catechin (Cat), hesperetin (H) and their equimolar binary mixtures were evaluated by means of cyclic voltammetry and luminol Co(II)–EDTA chemiluminescence, respectively, with a view to the prediction of a potential prooxidant or synergistic behaviour of mixtures of antioxidants. An important synergistic oxidation between Q and Cat was observed. The mixture CA/AA manifested the most prooxidant tendency among all the tested compounds and their mixtures. AA increased the antioxidant activity of Q and decreased that of Cat. Q and Cat regenerated CA, the prooxidant effect of the mixtures increasing with less e-delocalisation. Cat and CA have an antagonistic effect. H has the closest oxidation potential to CA and a slight increase of its antioxidant activity was registered when mixed with CA, which was not the case when it was combined with the other compounds or measured alone.

Effect of antioxidant mixtures on growth and ochratoxin a production of Aspergillus section Nigri species under different water activity conditions on peanut meal extract agar.

The effect of mixtures of antioxidants butylated hydroxyanisol (BHA) and propyl paraben (PP) on lag phase, growth rate and ochratoxin A (OTA) production by four Aspergillus section Nigri strains was evaluated on peanut meal extract agar (PMEA) under different water activities (aw). The antioxidant mixtures used were: BHA + PP (mM), M1 (0.5 + 0.5), M2 (1.0 + 0.5), M3 (2.5 + 0.5), M4 (0.5 + 1.0), M5 (1.0 + 1.0), M6 (2.5 + 1.0), M7 (5.0 + 2.5) and M8 (10 + 2.5). The mixture M8 completely suppressed mycelial growth for all strains. A significant stimulation in OTA production was observed with mixtures M1 to M5 mainly at the highest aw; whereas M6, M7 and M8 completely inhibited OTA production in all strains assayed; except M6 in A. carbonarius strain (RCP G). These results could enable a future intervention strategy to minimize OTA contamination.

Oxidative stress and bone markers in plasma of patients with long-bone fixative surgery: Role of antioxidants

It is well known that bone markers (e.g. osteocalcin and alkaline phosphatase) play a significant role in healing of bone fractures, whereas oxidative stress delay such healing. The present study aimed to investigate the effect of a mixture of antioxidants (vitamins A, C, E, and selenium) on oxidative stress parameters, and the levels of bone healing markers in the plasma of male patients following fixative surgery of long bones. Antioxidant tablets (300 µg vitamin A, 10 mg vitamin E, 60 mg vitamin C, and 75 µg selenium) were administered to groups 3 and 4 (10 patients in each) for 1 and 2 weeks, respectively, in addition to the regular postoperative treatment. Groups 1 (25 patients) and 2 (10 patients) received the regular post-operative treatment consisting of intravenous (I.V.) second generation of cephalosporin 1000 mg/day for 3 days, oral diclofenac 50 mg, and paracetamol 500 mg twice daily for 15 days. Osteocalcin level and alkaline phosphatase activity as well as antioxidant enzymes superoxide dismutase (SOD), glutathione reductase (GR), as well as glutathione (GSH), and thiobarbituric acid reactive substances (TBARS) as indices of oxidative stress, were determined in the plasma of all patients after 1 or 2 weeks of long-bone fixative surgery. The results revealed that osteocalcin level and the activity of alkaline phosphatase were markedly increased in the plasma of patients who received antioxidants for 2 weeks. In addition, after 1 and/or 2 weeks, the levels of TBARS were significantly lower in the antioxidant-treated patients compared with those who did not receive antioxidants. On the other hand, the activities of SOD and GR were markedly elevated in plasma of patients who received antioxidants after 1 or 2 weeks compared with patients who received regular therapy. Moreover, the level of plasma GSH was markedly increased only after 2 weeks in patients who received antioxidants. It is concluded that administration of antioxidant vitamins A, E, and C in addition to selenium could accelerate bone healing after long-bone fixative surgery. Therefore, antioxidants should be considered in designing therapeutic protocols in post-operative bone surgery.